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Biomedical subjects

M Sillis

Publications and source records attributed to M Sillis.

At least 19 recordsLinked to original sources

Microbiological profile of community-acquired pneumonia in adults over the last 20 years.

OBJECTIVES: To assess any change in the microbiological profile of community-acquired pneumonia (CAP) in our region over the last 20 years. METHODS: We compared hospital admissions aged between 15 and 74 (n = 61) in Norfolk (UK) for CAP over a 19-month period in 1982-3 (ST1) with all admissions aged over 16 (n = 99) over a 14-month period in 1999-2000 (ST2). Data were collected for ST1 as part of a prospective multicentred research study, in a period of high Mycoplasma pneumoniae activity. ST2 was a prospective study of clinical practice. Chlamydophila species were differentiated in ST2 using whole-cell immunofluorescence. RESULTS: A microbiological diagnosis was made in 38 (62%) in ST1 compared with 48 (48%) in ST2. Streptococcus pneumoniae remained the most common pathogen (26% in ST1, 25% in ST2). The incidence of M. pneumoniae was 18% in ST1 and 4% in ST2. The proportion of viral pathogens identified was similar: nine (15%) in ST1 and 14 (14%) in ST2. No cases of Chlamydophila pneumoniae were diagnosed in ST2. CONCLUSIONS: The microbiological profile of CAP in Norfolk (UK) has not changed over the last 20 years and C. pneumoniae is not a frequent pathogen.

Adolescent↗

Mental health of British farmers.

AIMS: To estimate the prevalence of neurotic symptoms in a sample of British farmers, to investigate whether farming characteristics are associated with psychiatric morbidity, and to test the hypothesis that British farmers have a higher prevalence of depression and thoughts of life not worth living than the British household population. METHODS: A total of 425 farmers from Hereford, Norwich, and Preston completed the Revised Clinical Interview Schedule (CIS-R) by computer between March and July 1999. The comparison cohort consisted of 9830 private householders aged 16-64 from the Office of Population Censuses and Surveys National Psychiatric Morbidity Surveys of Great Britain carried out in 1993 in which the CIS-R was administered. All analyses used the commands developed specifically for survey data available in Stata version 6.0. RESULTS: Taking a threshold of an overall score of 12 or more on the CIS-R, only 6% of farmers reported clinically relevant psychiatric morbidity. Psychiatric morbidity was not significantly associated with farm type or size in this study. Farmers reported a lower prevalence of psychiatric morbidity than the general population but were more likely to report thinking that life is not worth living, particularly after the low prevalence of psychiatric morbidity had been taken into account (odds ratio 2.56, 95% CI 1.39 to 4.69). When restricting the comparison to only rural or semirural householders, this increased risk was even more pronounced (odds ratio 3.26, 95% CI 1.51 to 7.02). CONCLUSIONS: The relation between depression and suicidal ideation seems to be quite different among farmers and the general population and warrants further investigation. We have shown it is possible to measure mental health systematically in a sample of British farmers. This study should be repeated in the aftermath of the foot and mouth crisis.

Adolescent↗

Simple method for determining biovar and serovar types of Ureaplasma urealyticum clinical isolates using PCR-single-strand conformation polymorphism analysis.

Ureaplasma urealyticum has been associated with urethritis in men, obstetric problems in women, and respiratory distress syndrome in preterm infants. U. urealyticum can be divided into two biovars comprising 14 serovars. Partial sequences of genes encoding the multiple-banded antigens of the cell surface are known. Using a commercially available precast DNA mutation detection gel system, we have developed a simple and reproducible PCR-single-strand conformation polymorphism analysis method for differentiating the biovars of this species that reveals five patterns among the 14 serovars and enables clinical isolates to be typed directly from broth cultures.

Bacterial Typing Techniques↗

Evidence of persisting serum antibodies to Escherichia coli O157 lipopolysaccharide and Verocytotoxin in members of rural communities in England.

The techniques of enzyme-linked immunosorbent assay (ELISA) and immunoblotting were used to examine a total of 1667 sera, from apparently healthy members of rural communities in England, for antibodies to the lipopolysaccharide (LPS) of Escherichia coli O157 and Verocytotoxins (VT). Twenty-nine sera from 22 individuals were shown to have antibodies specific for E. coli O157 LPS. Some of these lived on livestock farms and had occupational contact with cattle, suggesting that personnel working with farm animals may produce serum antibodies to the O157 LPS antigens. Fifteen people had IgG class antibodies to O157 LPS, suggesting long-term exposure to E. coli O157 and five people had serum antibodies on more than one occasion showing evidence of persistent antibodies to O157 LPS. Thirteen sera from 12 of 22 individuals also contained antibodies to VT1, VT2 or both toxins. Ten sera contained antibodies to VT1 and VT2, three sera contained antibodies to VT2 only.

Adult↗

Orf (contagious pustular dermatitis) in farmworkers: prevalence and risk factors in three areas of England.

Orf is a zoonotic skin disease which is commonly self-diagnosed by those who tend sheep and goats. This paper reports the prevalence, incidence and risk factors associated with the infection in a cohort of farmworkers from three areas of England, derived from the results of self-reporting and serology. Twenty-three per cent of those employed or living on a sheep farm reported ever having had orf, and the antibody serological profiles indicated a prevalence of 4 per cent and an annual incidence of 2.8 per cen. The main risk factors associated with the infection were contact with sheep, the size of the sheep flock, and contact with dogs.

Animal Husbandry↗

Multiplex polymerase chain reaction for the simultaneous detection of Mycoplasma pneumoniae, Chlamydia pneumoniae, and Chlamydia psittaci in respiratory samples.

AIMS: To develop a multiplex polymerase chain reaction (PCR) for the simultaneous detection of Mycoplasma pneumoniae, Chlamydia pneumoniae, and Chlamydia psittaci in respiratory samples. METHODS: Oligonucleotide primers for the amplification of the DNA of these three organisms were optimised for use in combination in the same reaction. PCR products were detected by hybridisation with pooled internal probes using an enzyme linked immunosorbent assay. Those with positive signals were further differentiated using species specific probes. Quality of DNA extraction and PCR inhibition were controlled by amplification of a human mitochondrial gene. A panel of 53 respiratory samples with known results was evaluated blindly. This was followed by a retrospective study on sputa collected from 244 patients with suspected community acquired pneumonia. RESULTS: The multiplex assay had a lower sensitivity than PCR with individual primers by about one log. The resultant sensitivity was considered acceptable for diagnostic use. Of the panel of 53 samples, nine of 11 M pneumoniae, 11 of 11 C pneumoniae, six of seven C psittaci, and 24 of 24 negative samples were correctly identified. Of the 244 patients with pneumonia, seven (2.9%) had detectable M pneumoniae, six (2.5%) had C pneumoniae, and one (0.4%) had C psittaci. The case notes from 11 patients were studied. The PCR finding was of possible significance in at least eight of these patients. CONCLUSIONS: This multiplex PCR assay has the potential to be used as a diagnostic and epidemiological tool. Further prospective studies are needed to establish its clinical value.

Adolescent↗

Low rates of ehrlichiosis and Lyme borreliosis in English farmworkers.

To determine the occupational significance of tick-borne zoonoses we sought serological evidence of Lyme borreliosis, human monocytic ehrlichiosis (HME) and human granulocytic ehrlichiosis (HGE) in a representative sample of farmworkers. Although around 20% reported ticks on their domestic and companion animals, few (< 2% per year) reported being bitten by ticks. Seroprevalence of Lyme borreliosis (0.2%), HME (0.2%) and HGE (1.5%) was low. Those seropositive for HGE were no more likely to report tick bites nor more likely to report ticks on their animals. This study provides evidence that farmworkers in England are exposed to tick-borne zoonoses but that they are uncommon. Since the severity of these diseases is linked to delays in diagnosis and treatment, clinicians should be aware of these diagnoses in patients from rural communities, with or without a self-reported history of tick bite.

Adult↗

Anticomplementary activity in serum samples from patients with acute parvovirus B19 infection.

Of 65 serum samples submitted for diagnostic purposes which proved to be anti-complementary by complement fixation test, 49 were parvovirus B19 IgM positive. Forty four of the 49 serum samples were from patients with arthropathy. Acute parvovirus B19 infection should be suspected when a patient has symptoms of disease of the joints and the serum is anticomplementary.

Acute Disease↗

Neurological disease associated with Mycoplasma pneumoniae infection. PCR evidence against a direct invasive mechanism.

Aims-To investigate the pathology in patients presenting with sudden onset neurological illnesses associated with Mycoplasma pneumoniae infection.Methods-M pneumoniae infection was diagnosed by a highly rigorous interpretation of serological markers initially using complement fixation, agglutination and IgM antibodies. Confirmation of the serological diagnosis was achieved using indirect immunofluorescence for IgM, IgA, and IgG. Serum and cerebrospinal fluid (CSF) samples from these patients were examined using the polymerase chain reaction to look for evidence of M pneumoniae DNA.Results-No M pneumoniae DNA was found in any serum or CSF samples. Diagnosis of M pneumoniae infection by agglutination and complement fixation antibodies was not always confirmed by indirect immunofluorescence.Conclusion-The neurological lesions in these patients do not appear to be caused by the direct invasion of M pneumoniae into the nervous system. The lesions may be an immune response to infection. Serological diagnosis of M pneumoniae continues to be a laboratory problem.

Journal Article↗

Direct sample polymerase chain reaction for the detection of Mycoplasma pneumoniae: a simple system for clinical application.

A semi-nested polymerase chain reaction system with primers derived from the P1 adhesin gene of Mycoplasma pneumoniae was evaluated for sensitivity and specificity for detection of M. pneumoniae. The method used target DNA within samples of M. pneumoniae broth cultures and clinical material, without a formal extraction process. The sensitivity for detection of DNA was found to be to a level of one copy per sample and was specific to M. pneumoniae only, discriminating from the other human mycoplasma and ureaplasma species tested. The system offers the opportunity for a simple and highly specific laboratory method for diagnosis which may be compared to currently available serological methods, and may provide another method of studying mycoplasma-induced pathology.

Base Sequence↗

Detection of Chlamydia pneumoniae and Chlamydia psittaci in sputum samples by PCR.

AIMS: To use the polymerase chain reaction (PCR) to detect Chlamydia pneumoniae and Chlamydia psittaci in sputum samples. METHODS: A nested PCR was developed, the first stage of which amplified DNA from both C pneumoniae and C psittaci while the second stage targeted specifically at C pneumoniae, allowing the two species to be differentiated. The primers were designed not to amplify sequences from C trachomatis. A panel of 26 sputum samples from patients with community acquired pneumonia evaluated previously by enzyme linked immunosorbent assay (ELISA), direct immunofluorescence (DIF), and culture was tested blind by PCR. Most of these specimens also had accompanying serial serum samples which were tested for species specific antibodies using microimmunofluorescence (micro-IF). RESULTS: PCR detected C pneumoniae DNA in 10 of the 26 samples and C psittaci DNA in four. There was good concordance between ELISA, DIF, micro-IF and PCR in the C pneumoniae group. Two of the C psittaci identified by PCR were labelled C pneumoniae by DIF but the PCR results were supported by serology or a history of bird contact. Of the PCR negative group: six were true negative results; two contained C trachomatis. There were four discrepant results. CONCLUSIONS: The data suggest that PCR is effective in the detection of C pneumoniae. The sensitivity for C psittaci is inevitably lower due to the strategy taken but specificity seemed to be good.

Bacterial Outer Membrane Proteins↗

Genital mycoplasmas revisited--an evaluation of a new culture medium.

The role of genital mycoplasmas as pathogens causing severe neonatal respiratory and central nervous system disease has been highlighted recently following publication of new data. These organisms are generally neglected by diagnostic laboratories in the United Kingdom, possibly due to the lack of a suitable commercially-available culture medium. We have evaluated the bioMerieux Mycoplasma-Lyo system using qualitative and quantitative studies and have found it to have suitable qualities and practical advantages for detection of Ureaplasma urealyticum and Mycoplasma hominis.

Culture Media↗

The differentiation of Chlamydia species by antigen detection in sputum specimens from patients with community-acquired acute respiratory infections.

An amplified enzyme immunoassay (IDEIA III: Dako Diagnostics Ltd) for detecting genus-specific chlamydia antigen was evaluated prospectively on 286 respiratory specimens from 275 patients presenting with community-acquired pneumonia or persistent chest infection. Nineteen patients had evidence of recent chlamydial infection, having two or more positive sputum or serological markers. Sputa from two other patients were ELISA-positive in the absence of other positive criteria and were regarded as false-positive results. When compared with a direct immunofluorescence test for chlamydial elementary bodies (EBs) using a genus-specific monoclonal antibody, the ELISA gave a positive predictive value of 91% and a negative predictive value of 99%. Non-specific problems with a wide variety of other micro-organisms isolated from the sputa were not encountered. Attempts to differentiate between Chlamydia psittaci, Chlamydia pneumoniae and Chlamydia trachomatis using genus-specific lipopolysaccharide reactive--and species-specific major outer membrane protein--monoclonal antibodies were encouraging and results were substantiated, in most patients, by the species-specific serological assays of the whole-cell-inclusion immunofluorescence or micro-immunofluorescence assays. The study demonstrated that antigen detection techniques offer scope for routine laboratories to diagnose chlamydial respiratory infections rapidly and reliably and may enable differentiation to species level. Although immunofluorescence offers marginally greater sensitivity and specificity when compared with ELISA, the latter is less subjective and less demanding. Sixty-eight per cent of these infections would have remained undiagnosed despite the general availability of ELISA tests.

Adult↗